3D Nephrotoxicity applications using an in vitro kidney proximal tubule cell based model on a microphysiological system.

Nortis’s microfluidic organ-on-chip technology

Date: Thursday, July 25, 2017

Time: 10:00 AM CST

Duration: 120 minutes (60 minutes each)

What Do They Cover?

Webinar 1:
The kidney proximal tubule is the primary site of drug-induced nephrotoxicity. I will describe the development of a 3-dimensional flow-directed proximal tubule microphysiological system (MPS). The kidney MPS recapitulates the synthetic, metabolic and transport activities of kidney proximal tubule cells. This MPS is as an ideal platform for ex vivo modeling of nephrotoxicity. Towards this goal, we have evaluated nephrotoxicity in response to challenge with multiple toxicants, including the heavy metal pollutant cadmium, antisense oligonucleotides, the antibiotic polymyxin B and the Chinese herbal product aristolochic acid. We believe that MPS technologies will have major impacts on predictive toxicity testing and human risk assessment. Animal and in vitro systems do not always faithfully recapitulate drug and xenobiotic responses in the clinic or occupational/environmental exposures, respectively. MPS technologies will refine safety assessment and reduce our need for surrogate animal testing. An ultimate goal is to create integrated human MPS organ systems that could replace animal models.

Webinar 2:
Nortis has developed a technology that is used to recapitulate functional units of human organs in microfluidic devices (chips). Such organ models include vasculature, kidney, and liver models for toxicology studies, blood-brain barrier models for drug transport studies, and vascularized tumor microenvironment models for drug efficacy studies. These models exhibit in-vivo like barrier function, transporter polarity, and enzymatic activity. Common architecture to these and other tissue models in the Nortis system is a 5 mm long, 0.125-mm diameter lumen lined with endothelial or epithelial cells that is completely surrounded by a 3D extracellular matrix, usually collagen I, and is continuously perfused with nutrient medium. Co-culture of cells can be compartmentalized by seeding cells into the extracellular matrix prior to polymerization, thus representing tissue parenchyma, and cells seeded into the tubular void to form endothelial microvessels or epithelial tubules traversing the pseudo parenchyma. By applying growth factor gradients, endothelial microvessels can be coaxed into sprouting vascular networks that interact with cells embedded in the extracellular matrix, thus building vascularized microenvironments. Experiments typically last seven to ten days, however they can last >40 days without loss of viability or function. The chips enable real-time microscopy during the course of the experiment and are compatible with quantitative metabolic reporter assays as well as live-cell and fixed-cell immunocytochemical staining. Effluent can be collected from lumenal and ablumenal compartments and assayed for metabolite and biomarker levels. Cells can be isolated for further analysis, including protein and mRNA profiling. Intact tubules can be excised and subjected to transmission electron microscopy or immunohistochemical analysis.

The Nortis microfluidic system is composed of disposable microfluidic chips, well-plate sized microscope-compatible perfusion platforms that secure the chips and house fluid reservoirs, and an incubator gas pump, which pressurizes air from inside the incubator to pneumatically drive fluid through the chips. Each chip allows three simultaneous experiments. With the current hardware setup, 72 independent experiments can be run simultaneously per incubator. In addition to selling perfusion hardware and chips, Nortis runs in-house contract service for tissue model development and drug testing for pharmaceutical and biotech companies. The data from these studies are showing remarkable consistency from experiment to experiment and in-vivo like results in terms of biomarker response, mRNA profiling, permeability, viability, and transporter function.

What Will You Learn?

3D Tissue Models

Kidney Proximal Tubule model

Organ-on-a Chip Technology

Drug Discovery & Safety Testing

Who Should Attend?

Academic and Pharma researchers

Speaker Bio

Edward Kelly, PhD

Edward Kelly earned his Ph.D. in Biochemistry from the University of Washington in the laboratory of Dr. Richard Palmiter, developing transgenic and knockout mouse models to study the function of the metal-binding protein metallothionein. Following a post-doctoral fellowship in molecular toxicology at the Department of Environmental Health with Dave Eaton, he ventured into Biotech, managing the Preclinical Bioanalytics group at Targeted Genetics Corporation, evaluating the safety and efficacy of gene therapies for diseases such as cystic fibrosis, rheumatoid arthritis and hemophilias. Upon his return to academia, his research interests have stayed within the realm of preclinical biology. His lab works on developing novel models to study normal human physiology and disease states, with a particular focus on cytochrome P450 enzymes and their role in endobiotic/xenobiotic metabolism. Specific areas include studying the heritable ocular disease, Bietti’s Crystalline Dystrophy and development of a human kidney “organ-on-a-chip”. Currently, he holds the position of Associate Professor in the Department of Pharmaceutics, Adjunct Associate Professor in the Department of Environmental and Occupational Health Sciences, and also serves as Co-Director of the Pharmaceutical Bioengineering Extension Program.

Speaker Bio

Henning Mann, PhD

Dr. Mann earned his PhD in physiological chemistry studying protein-protein and cell-protein interactions in the extracellular matrix. As a postdoctoral fellow he worked at the Fred Hutchinson Cancer Research Center in Seattle, WA, where he focused on understanding the expression, function and interactions of the immunostimulatory receptor-ligand pairs MIC and NKG2D. He then worked as Senior Staff Scientist at Seattle Biomed on developing and understanding in vitro culture assays for Plasmodium falciparum in perfused extracellular matrix microenvironments before joining Nortis, Inc. in 2012. As a Senior Research Scientist at Nortis, Dr. Mann leads the company's angiogenesis, vascularized tumor and immuno-oncology efforts.